氢化酶固定化研究进展

doi:10.12211/2096-8280.2024-022

摘要/Abstract

摘要：

氢化酶催化氢气向质子和电子的可逆转化，具有广阔的工业应用前景。但游离的氢化酶存在着对氧气敏感、传递电子速率慢等缺点。本文综述了碳材料、金属及半导体、高分子和金属-有机框架材料（MOFs）固定化氢化酶。碳材料具有价格低廉、比表面积大等优势。金属及半导体有着良好的导电性能和优异的催化性能。高分子材料具有良好的生物相容性和机械性能，可以提高氢化酶的稳定性和对氧气的耐受性。MOFs比表面积大，可设计调控，为理化性质不同的氢化酶提供了广泛的载体选择。复合材料固定化氢化酶可以结合不同材料的优势，拓宽固定化氢化酶的应用场景。固定化氢化酶可用于氢气的高效生产与应用以及生物不对称加氢制备手性化合物，为转变能源结构，实现绿色转型、解决环境问题提供了可选方案。

关键词: 氢化酶固定化, 生物电催化, 碳材料, 半导体材料, 高分子材料, 金属-有机框架（MOFs）

Abstract:

Hydrogenases catalyze the reversible conversion of hydrogen gas into protons and electrons which is promising for industrial application. However, free hydrogenases face challenges such as oxygen sensitivity and low electron transfer rates. This review summarized the immobilization of hydrogenases by carbon materials, metals, semiconductors, polymers and metal-organic-frameworks (MOFs). Carbon materials provide the advantages of low cost and large specific surface areas, while they tend to agglomerate. Hydrogenases are immobilizated on carbon materials through adsorption, usually involving electrostatic interactions and hydrophobic interactions, and are used in bioelectrocatalysis, biofuel cells, bioreactors et al. Metals and semiconductors, known for high conductivity and excellent reactive activity, are expensive and less stable. Through adsorption involving electrostatic interaction and hydrophobic interaction, immobilization of hydrogenases on metals and semiconductors are normally applied in bioelectrocatalysis, biofuel cells, photoelectrocatalysis et al. Polymers have good biocompatibility and mechanical strength but low conductivity. Immobilization of hydrogenases on polymers can improve the stability and oxygen tolerance of hydrogenases. Immobilization on polymers is realized through adsorption and entrapment, involving hydrogen bonds, hydrophobic interactions and π-π interactions, and is often used in bioelectrocatalysis, photoelectrocatalysis et al. MOFs are designable and have high specific surface areas, which provide wide choices for hydrogenases immobilization. However, MOFs tend to collapse in harsh conditions. Immobilization on MOFs through adsorption and entrapment involves coordinate bonds, hydrophobic interaction, and π-π interaction. Furthermore, the prospect of immobilization of hydrogenases by novel hybrid materials was proposed which can expand the applications of immobilized hydrogenases. Immobilization of hydrogenases facilitates the stability of hydrogenases, which can be applied in efficient production and application of hydrogen, as well as biological asymmetric hydrogenation for chiral medicine preparation. Immobilization of hydrogenases provide alternative options for transforming energy structures, realizing green manufacturing and solving environmental problems.

Key words: immobilization of hydrogenases, bioelectrocatalysis, carbon materials, semiconductors, polymers, metal-organic frameworks

中图分类号:

Q814.2

雷航彬, 何宁, 李斐煊, 董玲玲, 王世珍. 氢化酶固定化研究进展[J]. 合成生物学, DOI: 10.12211/2096-8280.2024-022.

Hangbin LEI, Ning HE, Feixuan LI, Lingling DONG, Shizhen WANG. Advance of the immobilization of hydrogenases[J]. Synthetic Biology Journal, DOI: 10.12211/2096-8280.2024-022.

图/表 7

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材料	具体固定化载体	氢化酶来源	分类	应用	参考文献
碳材料	石墨	Escherichia coli ( [NiFe])	[NiFe]	生物电催化	[18]
	石墨	Aquifex aeolicus	[NiFe]	生物电催化	[19]
	石墨	Clostridium acetobutylicum	[FeFe]	生物电催化	[20]
	石墨	Ralstonia eutropha	[NiFe]	生物电催化	[21]
	石墨	Desulfovibrio gigas	[NiFe]	生物电催化	[22]
	石墨	Ralstonia metallidurans	[NiFe]	生物燃料电池	[23]
	石墨	Allochromatium vinosum	[NiFe]	动力学研究	[24]
	碳黑	Ralstonia eutropha	[NiFe]	光谱电化学研究	[25]
	碳丝	Thiocapsa roseopersicina	[NiFe]	生物燃料电池	[26]
	碳丝	Thiocapsa roseopersicina	[NiFe]	生物反应器	[27]
	碳纸	Pyrococcus furiosus	[NiFe]	生物燃料电池	[28]
	碳毡	Clostridium acetobutylicum	[NiFe]	生物燃料电池	[29]
	单壁碳纳米管	Clostridium acetobutylicum	[FeFe]	生物电催化	[30]
	单壁碳纳米管	Clostridium acetobutylicum	[FeFe]	生物电催化	[31]
	单壁碳纳米管	Allochromatium vinosum	[FeFe]	生物电催化	[32]
	多壁碳纳米管	Aquifex aeolicus	[NiFe]	电化学传感器	[33]
	多壁碳纳米管	Ralstonia eutropha Aquifex aeolicus	[NiFe]	生物燃料电池	[34]

材料	具体固定化载体	氢化酶来源	分类	应用	参考文献
碳材料	石墨	Escherichia coli ( [NiFe])	[NiFe]	生物电催化	[18]
	石墨	Aquifex aeolicus	[NiFe]	生物电催化	[19]
	石墨	Clostridium acetobutylicum	[FeFe]	生物电催化	[20]
	石墨	Ralstonia eutropha	[NiFe]	生物电催化	[21]
	石墨	Desulfovibrio gigas	[NiFe]	生物电催化	[22]
	石墨	Ralstonia metallidurans	[NiFe]	生物燃料电池	[23]
	石墨	Allochromatium vinosum	[NiFe]	动力学研究	[24]
	碳黑	Ralstonia eutropha	[NiFe]	光谱电化学研究	[25]
	碳丝	Thiocapsa roseopersicina	[NiFe]	生物燃料电池	[26]
	碳丝	Thiocapsa roseopersicina	[NiFe]	生物反应器	[27]
	碳纸	Pyrococcus furiosus	[NiFe]	生物燃料电池	[28]
	碳毡	Clostridium acetobutylicum	[NiFe]	生物燃料电池	[29]
	单壁碳纳米管	Clostridium acetobutylicum	[FeFe]	生物电催化	[30]
	单壁碳纳米管	Clostridium acetobutylicum	[FeFe]	生物电催化	[31]
	单壁碳纳米管	Allochromatium vinosum	[FeFe]	生物电催化	[32]
	多壁碳纳米管	Aquifex aeolicus	[NiFe]	电化学传感器	[33]
	多壁碳纳米管	Ralstonia eutropha Aquifex aeolicus	[NiFe]	生物燃料电池	[34]

材料	具体固定化载体	氢化酶来源	分类	应用	参考文献
金属	金电极	Chlamydomonas reinhardtii	[FeFe]	生物电催化	[43]
	金电极	Desulfovibrio vulgaris	[NiFe]	电化学研究	[44]
	金电极	Desulfovibrio vulgaris	[NiFe]	电化学分析	[45]
	金电极	Ralstonia eutropha	[NiFe]	电化学研究	[46]
	金电极	Desulfovibrio vulgaris	[NiFe]	生物电催化	[47]
	金电极	Ralstonia eutropha	[NiFe]	生物电催化	[48]
	硫醇修饰金电极	Aquifex aeolicus	[NiFe]	生物电催化	[49]
	紫精修饰金电极	Desulfovibrio desulfuricans	[FeFe]	生物燃料电池	[50]
	碳纳米管修饰金电极	Desulfovibrio gigas	[NiFe]	生物燃料电池	[51]
	碳纳米管修饰金电极	Desulfovibrio fructosovorans	[NiFe]	生物燃料电池	[52]
	纳米金电极	Aquifex aeolicus	[NiFe]	生物燃料电池	[53]
	纳米金电极	Allochromatium vinosum	[NiFe]	单酶分子电化学	[54]
	银纳米团簇	Escherichia coli	[NiFe]	光电催化	[55]
半导体	TiO₂	Thiocapsa roseopersicina	[NiFe]	光电催化	[56]
	TiO₂	Desulfomicrobium baculatum	[NiFeSe]	光电催化	[57]
	TiO₂	Desulfomicrobium baculatum	[NiFeSe]	光电催化	[58]
	TiO₂	Clostridium acetobutylicum	[FeFe]	生物电催化	[59]
	PVK\|IO-TiO₂	Desulfovibrio vulgaris	[NiFeSe]	光电化学集成系统	[60]
	ITO	Desulfovibrio vulgaris	[NiFe]	光电催化	[61]
	ITO	Desulfomicrobium baculatum	[NiFeSe]	光电催化	[62]
	ITO	Ralstonia eutropha	[NiFe]	生物电子设备	[63]
	CN_x(氮化碳)	Desulfomicrobium baculatum	[NiFeSe]	光电催化	[64]
	CdS	Clostridium acetobutylicum	[FeFe]	光电催化	[65]
	CdS	Clostridium acetobutylicum	[FeFe]	电子转移动力学研究	[66]
	CdTe	Clostridium acetobutylicum	[FeFe]	光电催化	[67]
	CdTe	Thiocapsa roseopersicina	[NiFe]	光电催化	[68]
	In₂S₃	Desulfovibrio vulgaris	[NiFeSe]	光电催化	[69]
	FTO-NiO-In₂S₃	Desulfovibrio vulgaris	[NiFeSe]	光电催化	[70]

材料	具体固定化载体	氢化酶来源	分类	应用	参考文献
金属	金电极	Chlamydomonas reinhardtii	[FeFe]	生物电催化	[43]
	金电极	Desulfovibrio vulgaris	[NiFe]	电化学研究	[44]
	金电极	Desulfovibrio vulgaris	[NiFe]	电化学分析	[45]
	金电极	Ralstonia eutropha	[NiFe]	电化学研究	[46]
	金电极	Desulfovibrio vulgaris	[NiFe]	生物电催化	[47]
	金电极	Ralstonia eutropha	[NiFe]	生物电催化	[48]
	硫醇修饰金电极	Aquifex aeolicus	[NiFe]	生物电催化	[49]
	紫精修饰金电极	Desulfovibrio desulfuricans	[FeFe]	生物燃料电池	[50]
	碳纳米管修饰金电极	Desulfovibrio gigas	[NiFe]	生物燃料电池	[51]
	碳纳米管修饰金电极	Desulfovibrio fructosovorans	[NiFe]	生物燃料电池	[52]
	纳米金电极	Aquifex aeolicus	[NiFe]	生物燃料电池	[53]
	纳米金电极	Allochromatium vinosum	[NiFe]	单酶分子电化学	[54]
	银纳米团簇	Escherichia coli	[NiFe]	光电催化	[55]
半导体	TiO₂	Thiocapsa roseopersicina	[NiFe]	光电催化	[56]
	TiO₂	Desulfomicrobium baculatum	[NiFeSe]	光电催化	[57]
	TiO₂	Desulfomicrobium baculatum	[NiFeSe]	光电催化	[58]
	TiO₂	Clostridium acetobutylicum	[FeFe]	生物电催化	[59]
	PVK\|IO-TiO₂	Desulfovibrio vulgaris	[NiFeSe]	光电化学集成系统	[60]
	ITO	Desulfovibrio vulgaris	[NiFe]	光电催化	[61]
	ITO	Desulfomicrobium baculatum	[NiFeSe]	光电催化	[62]
	ITO	Ralstonia eutropha	[NiFe]	生物电子设备	[63]
	CN_x(氮化碳)	Desulfomicrobium baculatum	[NiFeSe]	光电催化	[64]
	CdS	Clostridium acetobutylicum	[FeFe]	光电催化	[65]
	CdS	Clostridium acetobutylicum	[FeFe]	电子转移动力学研究	[66]
	CdTe	Clostridium acetobutylicum	[FeFe]	光电催化	[67]
	CdTe	Thiocapsa roseopersicina	[NiFe]	光电催化	[68]
	In₂S₃	Desulfovibrio vulgaris	[NiFeSe]	光电催化	[69]
	FTO-NiO-In₂S₃	Desulfovibrio vulgaris	[NiFeSe]	光电催化	[70]

固定化载体	氢化酶来源	分类	储存/反应条件及剩余酶活	参考文献
紫精凝胶	Desulfovibrio vulgaris	[NiFe]	4 ℃，磷酸盐缓冲液pH=7.0，储存20天保持50％的酶活	[85]
聚合物多孔凝胶	Clostridium pasteurianum	[Fe]	室温，厌氧缓冲液pH=8.0，储存28天，保持70％的活性	[86]
聚合物多孔凝胶	Lamprobacter modestogalophilus	[NiFe]	室温，厌氧缓冲液pH=8.0，储存28天，保持50％的活性	[86]
海藻酸钙凝胶	Desulphovibrio desulphuricans	[NiFe]	4 ℃，Tris-HCl缓冲液pH=7.5，储存40天保持60％的活性	[87]
海藻酸钙凝胶	DesuEfouibrio sp.	[NiFe]	30 ℃，Tris-HCl缓冲液pH=7.6，反应50 h保持40％的活性	[88]
阴离子交换树脂	Ralstonia eutropha	[NiFe]	35 ℃，Tris-HCl缓冲液pH=8.0，反应32 h保持50％的活性	[89]
琼脂糖凝胶	Chromatium vinosum	[NiFe]	65 ℃，Tris-HCl缓冲液pH=8.0，孵育80 min保持50％的稳定性	[90]